MARVEL-ous Extracellular vesicles carry RXLR effectors into host plant cells

MARVEL-ous 细胞外囊泡携带 RXLR 效应子进入宿主植物细胞

• 批准号: BB/Y002067/1
• 负责人: Paul Birch
• 金额: $ 55.07万
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY002067%2F1
• 关键词: MARVEL; ous; Extracellular; vesicles; carry

In 2009, the World Summit on Food Security set a global challenge of 60% increase in crop production by 2050 to meet predicted demands for food. Damage from pests and pathogens causes losses of up to 23% of global production of the five most important food crops, representing a significant threat to food security. Oomycete plant pathogens cause severe damage to many major food crops. For example, the potato and tomato late blight pathogen Phytophthora infestans causes annual global losses estimated to exceed $10 bn. Plant pathogens such as P. infestans cause disease by delivering an arsenal of virulence proteins, called effectors, into plant tissues during infection. Amongst these are hundreds of so-called RXLR effectors that are delivered inside living plant cells where they often target host proteins to suppress immunity. A major scientific challenge in the plant-microbe interaction field is to understand how RXLR effectors are secreted and delivered into plant cells.We have made some key breakthroughs forming the basis for this proposal. We have generated data showing that RXLR effectors are secreted in association with membrane-bound extracellular vesicles (EVs). We have discovered other proteins associated with these EVs, including proteins with MARVEL domains containing multiple transmembrane helices. These MARVEL proteins co-localise with RXLR effectors in vesicles, and we hypothesise that they are novel markers of EVs associated with RXLR effector delivery. In our proposal we aim to confirm whether MARVEL proteins are markers of EVs associated with RXLRs and we will use cutting-edge super-resolution microscopy to provide a detailed and deep investigation of their secretion and delivery at the interface between P. infestans and its hosts. We will remove the MARVEL proteins by gene silencing and CRISPR/cas12 knockout techniques to determine their importance to effector secretion and thus to infection. Finally, we will tag the MARVEL proteins and use them to capture EVs following growth in media and during infection and study the EV proteome to identify factors involved in biogenesis, structure and function of EVs, and to determine the cargo of virulence proteins delivered to hosts during infection.This work will provide a deep understanding of a key fundamental effector delivery mechanism likely to be shared across oomycete plant pathogens which can be targeted by agrochemical or biotechnological means to prevent crop disease.

2009年，世界粮食安全峰会引起了全球挑战，即到2050年，农作物产量增加了60％，以满足预测的粮食需求。害虫和病原体的损害导致全球生产的23％损失五种最重要的粮食作物，这代表了对粮食安全的重大威胁。 Oomycete植物病原体对许多主要粮食作物造成严重损害。例如，马铃薯和番茄晚期病原体疫霉菌Infestans导致年度全球损失估计超过10亿美元。植物病原体（例如侵注疟原虫）通过在感染过程中将毒力蛋白（称为效应子）的武器递送到植物组织中，从而引起疾病。其中包括数百种所谓的RXLR效应子，它们在活植物细胞内传递，它们通常靶向宿主蛋白以抑制免疫力。植物菌相互作用领域的一个主要科学挑战是了解RXLR效应子是如何分泌并传递到植物细胞中的。我们做出了一些关键的突破，为该提案构成了基础。我们生成的数据表明，RXLR效应子与膜结合的细胞外囊泡（EV）分泌。我们发现了与这些EV相关的其他蛋白质，包括具有含有多个跨膜螺旋的奇迹结构域的蛋白质。这些奇迹蛋白与囊泡中的RXLR效应子共定位，我们假设它们是与RXLR效应子递送相关的EV的新颖标志。在我们的提案中，我们旨在确认奇迹蛋白是否是与RXLR相关的电动汽车的标志，我们将使用尖端的超分辨率显微镜来提供对它们在Infestans及其宿主之间界面的分泌和递送的详细和深入研究。我们将通过基因沉默和CRISPR/CAS12敲除技术去除奇迹蛋白，以确定它们对效应子分泌的重要性，从而确定其对感染的重要性。 Finally, we will tag the MARVEL proteins and use them to capture EVs following growth in media and during infection and study the EV proteome to identify factors involved in biogenesis, structure and function of EVs, and to determine the cargo of virulence proteins delivered to hosts during infection.This work will provide a deep understanding of a key fundamental effector delivery mechanism likely to be shared across oomycete plant pathogens which can be targeted by agrochemical or预防作物疾病的生物技术手段。